AIMTo determine the effects of azide methyl anthraquinone derivative (AMAD) on growth inhibition and inducing apoptosis of multidrug resistant (MDR) KBv200 cells and parental drug-sensitive KB cells.

METHODSCytotoxicity was determined by tetrazolium (MTF) assay. Reactive oxygen species (ROS) levels and mitochondrial membrane potential (deltapsi(m)) in cells were labeled with DCFH-DA and DiOC6 and tested by flow cytometry. Annexin V stain and DNA ladder were used to examine the apoptosis of KB and KBv200 cells induced by AMAD.

RESULTSAMAD was shown to inhibit the growth of KB and KBv200 cells significantly in a concentration-dependent manner, with mean IC50 of 0.36 and 0.45 micromol x L(-1), respectively. The generation of ROS increased obviously after the cells were treated with AMAD for 12 h, up to the peak in 24 h, meanwhile the levels of deltapsi(m) were time-dependently decreased. DNA fragmentation appeared on the agarose gel. Annexin V stain showed AMAD induced apoptosis of KB and KBv200 cells also in a concentration-dependent manner.

CONCLUSIONAMAD showed inhibitory effect on both MDR KBv200 cells and parental drug-sensitive KB cells. The mechanism of action was associated with the increase of the cellular ROS level and the decrease of the mitochondrial membrane potential induced by AMAD, which result in cell apoptosis.

Anthraquinones ; administration & dosage ; chemistry ; pharmacology ; Antineoplastic Agents ; administration & dosage ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; KB Cells ; Mitochondria ; physiology ; Molecular Structure ; Mouth Floor ; Mouth Neoplasms ; pathology ; Reactive Oxygen Species ; metabolism ; Vincristine ; pharmacology

Objective To help the medical student to master the principle and operation of magnetic resonance imaging instrument.Methods The imaging principle,structure and functions of HT-3DNMR-25 magnetic resonance imaging instrument were described,and its application to experiment teaching was introduced including one-dimension imaging,two-dimension imaging,free induction decay signal observation and etc.Results The instrument facilitated the student to master technical principle,imaging method as well as parameters setup and regulation,and could be a supplement to clinical practical teaching.Conclusion The instrument plays an irreplaceable role in teaching,and thus is worthy promoting in colleges.

OBJECTIVETo construct lentivector carrying Tie2-Small interfering RNA (SiRNA), so as to study its influence on malignant melanoma cells.

METHODSRecombinant plasmid pSilencer 1.0-U6-Tie2-siRNA and plasmid pNL-EGFP were digested with XbaI, ligated a target lentiviral transfer plasmid of pNL-EGFP-U6-Tie2-I or pNL-EGFP-U6-Tie2-II, and then the electrophoresis clones was sequenced. Plasmids of pNL-EGFP-U6-Tie2-I and pNL-EGFP-U6-Tie2-II were constructed and combined with pVSVG and pHelper, respectively, to constitute lentiviral vector system of three plasmids. The Lentiviral vector system was transfected into 293T cell to produce pNL-EGFP-U6-Tie2- I and pNL-EGFP-U6-Tie2-II lentivirus. Then the supernatant was collected to determine the titer. Malignant melanoma cells were infected by both lentiviruses and identified by Realtime RT-PCR to assess inhibitory efficiency.

RESULTSThe recombinant lentiviral vectors of Tie2-RNAi were constructed successfully which were analyzed with restriction enzyme digestion and identified by sequencing. And the titer of lentiviral vector was 8.8 x 10(3)/ml, which was determined by 293T cell. The results of Realtime RT-PCR demonstrated that the lentiviral vectors of Tie2-RNAi could infect malignant melanoma cells and inhibit the expression of Tie2 genes in malignant melanoma cells (P<0.01). There was no significant difference in the expression level (P>0.05) between the two lentiviral vectors of Tie2-RNAi.

CONCLUSIONSLentivector carrying Tie2-SiRNA can be constructed successfully and inhibit the expression of Tie2 gene in vitro significantly. The study will supply the theory basis for the further research on the inhibition of tumor growth in vivo.

Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Melanoma ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, TIE-2 ; genetics ; Transfection

OBJECTIVETo investigate the effects of simvastatin (Sim) and the interference by mevalonate (MVA) against its effect on DNA synthesis in rat cardiac fibroblasts (CFs).

METHODSCFs were isolated from neonatal SD rats by trypsin digestion and growth-arrested CFs were stimulated with Sim and/or MVA at varied concentrations for different time lengths, and the DNA synthesis in the cells was measured by (3)H-thymidine ((3)H-TdR) incorporation assay.

RESULTSSim decreased (3)H-TdR incorporation in the CFs in a concentration-dependent manner, and (3)H-TdR incorporation was significantly lower in cells treated with 1 x 10(-6) and 1 x 10(-5) mol/L Sim (1,175+/-202.66 and 771+/-164.86 cpm/2000 cells, respectively) than in the control cells (1,608+/-204.32 cpm/2000 cells, P<0.01). As the treatment time with 1 x 10(-5) mol/L Sim prolonged (for 6, 12, 18, 24, 36, 42, and 48 h), (3)H-TdR incorporation in CFs decreased gradually, showing an obvious inverse correlation with the treatment time (r=-919, P<0.01). (3)H-TdR incorporation in cells treated with 1 x 10(-6) to 1 x 10(-3) mol/L MVA and 1 x 10(-5) mol/L Sim rose steadily as MVA concentration increased. A significant difference in the incorporation was found between cells treated with both 1 x 10(-4)/1 x 10(-3) mol/L MVA and 1 x 10(-5) mol/L Sim (1,612+/-308.57 and 1,995+/-353.83 cpm/2000 cells, respectively) and the cells with 1 x 10(-5) mol/L Sim treatment alone (P<0.01); difference was also noted between cells treated with 1 x 10(-5) mol/L MVA and the control cells (P<0.05), but treatment with 1 x 10(-6) mol/L MVA did not produce much difference in comparison with the control cells (P>0.05) With the increase of treatment time (for 6, 12, 18, 24, 36, 42, 48 h), 1 x 10(-3) mol/L MVA caused steady increase in (3)H-TdR incorporation in the CFs, showing a significant positive correlation with the treatment time (r=0.968, P<0.01).

CONCLUSIONSim can decrease DNA synthesis in rat CFs and postpone the occurrence of myocardial fibrosis, which can be reversed by MVA.

Animals ; Animals, Newborn ; Cells, Cultured ; DNA ; biosynthesis ; Dose-Response Relationship, Drug ; Female ; Fibroblasts ; cytology ; drug effects ; metabolism ; Fibrosis ; prevention & control ; Hypolipidemic Agents ; pharmacology ; Male ; Mevalonic Acid ; pharmacology ; Myocardium ; metabolism ; pathology ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Simvastatin ; pharmacology ; Time Factors

Objective To study the variational features of the brainstem auditory evoked potential (BAEP) and the preservation of the facial and auditory nerves during the microsurgery in patients with acoustic neuromas (AN). Methods We retrospectively analyzed the data of 76 patients with AN, admitted to our hospital fi'om May 2006 to May 2009: preoperative BAEP were examined; the peak latency (PL) of Ⅰ, Ⅲ and Ⅴ waves and their interpeak latency (IPL) were noted and compared. The tumors were microsurgically removed under intraoperative electrophysiologic monitoring to preserve the facial and auditory nerves. The relationships between the preservation of facial and auditory nerves and both the tumor size and the utilization of intraoperative monitoring were discussed. Results Before the operation, the 100% abnormal rate of BAEP was showed on the diseased side with the disappearance of Ⅲ and Ⅴ waves as its main features, while 82.9% (63/76) of that was noted on the contralateral side with the IPL ratio of Ⅲ-Ⅴ / Ⅰ-Ⅲ greater than or equal to 1 as its main features; the BAEP features between the 2 sides had statistical significance (P<0.05). Seventy patients (92.1%) were found abnormal of BAEP on the contralateral side during the operation; the BAEP features of the contralateral side had statistical significance between preoperation and intraoperation (P<0.05). The tumors were totally removed in 69 (90.8%), subtotally removed in 6 (7.9%) and partially removed in 2 (1.3%). The preservation of facial and auditory nerves increased obviously through monitoring the response of BAEP during the surgery and a positive correlation was observed between the preservation of facial and auditory nerves and the response of BAEP in the contralateral side(r=0.912, P=0.000; r=0.613, P=0.000). However, an inverse correlation was observed between the preservation of facial and auditory nerves and the size of AN (r=-0.869, P=0.000; r=-0.738, P=0.000). Conclusion Preoperative BAEP can help to make an exact diagnosis on patients with AN and the preservation of facial and auditory nerves can increase obviously through monitoring the normal response of BAEP during the surgery. The anatomy preservation of facial and auditory nerves depends on the size of AN.

OBJECTIVETo study the clinical effect and mechanisms of specific sublingual immunotherapy (SLIT) for the treatment of allergic rhinitis or asthma in children.

METHODSThirty children suffering from Dermatophagoides farinae-allergic rhinitis or asthma (case group) and 30 healthy children (control group) were enrolled in this study. The case group accepted SLIT between January and December 2011. The ratio of forced expiratory volume in one second (FEV1) and its expected value, the ratio of airway resistance and its expected value, peripheral blood eosinophil (Eos) count and serum levels of IL-17 and IL-35 were measured before treatment and one and two years after treatment. The rhinitis or asthma symptom scores were rated and the level of asthma control was monitored.

RESULTSSerum IL-17 level in the case group was significantly higher than in the control group before treatment and one year after treatment (P<0.01). Furthermore, serum IL-17 level in the case group gradually decreased from before treatment to 1 year to 2 years after treatment (P<0.01). By two years of treatment, there was no significant difference in serum IL-17 level between the case and control groups (P>0.05). The changes of serum IL-35 level after treatment were opposite to serum IL-17 in the case group. The ratio of FEV1 and its expected value gradually increased from before treatment to 1 year to 2 years after treatment (P<0.01) in the case group. In contrast, the change of the ratio of airway resistance and its expected value and Eos count gradually decreased from before treatment to 1 year to 2 years after treatment (P<0.01) in the case group. More patients achieved improved rhinitis or asthma symptom scores two years after treatment than one year after treatment in the case group (P<0.01). SLIT was effective in 85% of children with allergic rhinitis one after treatment vs 100% two years after treatment. Asthma control was observed in 76% of the asthmatic patients one after treatment vs 92% two years after treatment.

CONCLUSIONSSLIT is effective for allergic rhinitis and asthma in children, and the treatment period of two years seems to be superior to one year. The mechanism of action of SLIT for the treatment of allergic rhinitis and asthma may be associated with inhibition of IL-17 expression and promotion of IL-35 expression.

Adolescent ; Asthma ; blood ; therapy ; Child ; Child, Preschool ; Female ; Humans ; Interleukin-17 ; blood ; Interleukins ; blood ; Male ; Rhinitis, Allergic ; blood ; therapy ; Sublingual Immunotherapy

BACKGROUNDData on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.

METHODSThe survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.

RESULTSThe analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).

CONCLUSIONSThe prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.

Adult ; Aged ; Awareness ; Female ; Humans ; Hypertension ; complications ; epidemiology ; therapy ; Male ; Middle Aged ; Prevalence ; Renal Insufficiency, Chronic ; complications